Selected article for: "expression vector and transient expression"
Author: Whitehouse, Caroline; Burchell, Joy; Gschmeissner, Stephen; Brockhausen, Inka; Lloyd, Kenneth O.; Taylor-Papadimitriou, Joyce
Title: A Transfected Sialyltransferase That Is Elevated in Breast Cancer and Localizes to the medial/trans-Golgi Apparatus Inhibits the Development of core-2–based O-Glycans
  • Document date: 1997_6_16
    • ID: pwgj97kz_8
    Snippet: The cDNA encoding human ␣ 2,3 SAT (O) was cloned into the HindIII/ XbaI site of pBluescript (Stratagene, La Jolla, CA). This was digested with MroI and XbaI, and two annealing oligos encoding the myc epitope were inserted. The sequence of these oligos was as follows: 5 Ј CCGGAT-CTTCAAGGGGAGACCTGAACAGAAACTGATCTCTGAAGAAG-ACCTGTGAT 5 Ј CTAGATCACAGGTCTTCTTCAGAGATCAGTTT-CTGTTCAGGTCTCCCCTTGAAGAT. Bases encoding the myc epitope recognized by the 9E1.....
    Document: The cDNA encoding human ␣ 2,3 SAT (O) was cloned into the HindIII/ XbaI site of pBluescript (Stratagene, La Jolla, CA). This was digested with MroI and XbaI, and two annealing oligos encoding the myc epitope were inserted. The sequence of these oligos was as follows: 5 Ј CCGGAT-CTTCAAGGGGAGACCTGAACAGAAACTGATCTCTGAAGAAG-ACCTGTGAT 5 Ј CTAGATCACAGGTCTTCTTCAGAGATCAGTTT-CTGTTCAGGTCTCCCCTTGAAGAT. Bases encoding the myc epitope recognized by the 9E10 mAb (Evan et al., 1985) are underlined. The product was digested with HindIII/NotI and cloned into the pcDNAIneo expression vector (Invitrogen, San Diego, CA). This neo3STMYC construct was sequenced, and expression of the myc epitope tag was checked by transient expression in COS cells before being used for stable transfection of T47D cells.

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